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J. Biol. Chem., Vol. 281, Issue 39, 28731-28736, September 29, 2006

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Identification and Characterization of the CD226 Gene Promoter^*

Jin-Long Jian, Can-Sheng Zhu, Zhu-Wei Xu, Wei-Ming Ouyang, Dong-Chu Ma, Yuan Zhang, Li-Jie Chen, An-Gang Yang, and Bo-Quan Jin¹

From the Department of Immunology, Fourth Military Medical University, Xi'an City, Shannxi Province 710032, China and Department of Experimental Medicine, Northern Hospital, Wenhua Road, Shenyang City, Liaoning Province 110015, China

CD226 is one of the main activating receptors on natural killer cells, and it can induce cytotoxicity to target cells through interaction with its ligands CD155 or CD112. CD226 is also involved in T cell differentiation, activation, and cytotoxicity. The expression of CD226 on natural killer cells and T cells can be regulated by cytokines and chemical stimuli; however, the mechanism of the regulation of the CD226 gene is still unknown. In this study, we have identified two promoters in the human CD226 gene named P1 and P2, which are located at –810 to –287 bp and +33 to +213 bp, respectively, and a negative regulation element between P1 and P2. Both P1 and P2 can be regulated by phorbol ester (12-O-tetradecanoylphorbol-13-acetate) and calcium ionophore (A23187 [GenBank] ). Bioinformatics analysis shows that, within this CD226 gene region, there are putative binding sites for transcription factors AP-1, Sp1, PEA3, and Ets-1. We have found that transcription factor activating protein-1 (AP-1) can up-regulate CD226 promoters P1 and P2 in human hepatocarcinoma cells, a hepatocarcinoma cell line with low expression of endogenous AP-1 and Ets-1. Interestingly, the transcription factor Ets-1 promotes AP-1-induced P2 activity but inhibits AP-1-induced P1 activity for which a 10-bp AP-1/Ets-1 composite site (CCTTCCTTCC) in P1 may be responsible.

Received for publication, February 24, 2006 , and in revised form, July 17, 2006.

^* This work was supported by the National Key Basic Research Program of China (Grant 2001CB510004) and National Natural Science Foundation of China (Grant 30030130). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Dept. of Immunology, Fourth Military Medical University, 17 Changle Rd., Xi'an 710032, China. Tel.: 86-29-8477-4598; Fax: 86-29-8325-3816; E-mail: immu_jin{at}fmmu.edu.cn.

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